SK287276B6 - Crystalline form of the methanesulfonic acid addition salt - Google Patents

Abstract

It is described a new crystalline form of the methanesulfonic acid addition salt of 4-(4-methylpiperazin-1- ylmethyl)-N-[4-methyl-3-(4-pyridin-3-yl)pyrimidin-2- ylamino)phenyl]-benzamide of formula (I), which may be used for example for tumour therapy.

Description

The present invention relates to a specific form of a methanesulfonic acid addition salt of 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) amino] phenyl} benzamide containing typical crystals .

BACKGROUND OF THE INVENTION

Preparation of 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl} benzamide and its use, in particular as an antitumor agent is described in Example 21 of EP-A-0 564 409, published on October 6, 1993 and co-pending applications in many other states. This compound is given in free form only (not as a salt) in the examples of the publications listed.

SUMMARY OF THE INVENTION

The invention relates to a crystalline form of a monomethanesulfonic acid addition salt with a compound of formula (I)

(I) characterized by a melting point of about 226 ° C, defined as the onset of melting in the differential scanning calorimetry diagram.

This crystalline form has acicular crystals.

In particular, the invention relates to a substantially pure crystalline form of the methanesulfonic acid addition salt of 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyridin-3-yl) pyrimidin-2- ylamino] phenyl} benzamide of formula (I).

Where the term methanesulfonic acid salt with a compound of formula (I) or 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3)] is used or will be used. -yl) pyrimidin-2-ylamino] phenyl) benzamide, in particular the methanesulfonic acid salt of formula (II)

(Π).

By "substantially pure" in the context of the present invention is meant that in particular at least 90, preferably 95 and most preferably at least 99 wt. the acid addition salt crystals of formula (I) is contained in the crystalline form of the invention.

Accordingly, the invention particularly relates to such methanesulfonic acid addition salt forms of a compound of formula (I) in which the crystals of the crystalline form of the invention are contained in substantially pure form together with other crystalline forms and / or amorphous forms of 4- (4-methylpiperazine) methanesulfonate. -l-ylmethyl) -N- {4-methyl [3- (4-pyridin-3-yl) -pyrimidin-2-ylamino] phenyl} benzamide.

The new crystalline form has the following characteristics:

The melting point of the α-crystalline form is 226 ° C (start of melting).

α-crystalline form of methanesulfonic acid addition salt of 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl} benzamide obtained, for example, by precipitation of this salt from solution in a solvent other than an alcohol such as methanol and without the addition of seed crystals of the β-crystalline form.

The conditions for the selective preparation of the individual crystalline forms are not critical. In general, for example, it is possible to vary parameters such as the weight ratio of the methanesulfonic acid addition salt of the compound of formula (I) to the solvent.

The fact is that the α-crystalline form of the methanesulfonic acid addition salt of the compound of formula (I) is metastable at room temperature.

Obviously, at 25 ° C, the α-crystalline form is hygroscopic and rapidly absorbs water, so that at 93% relative humidity, the sample has some amorphous form, the α-crystalline form liquefies at 97% relative humidity.

Methanesulfonic acid addition salt of the compound of formula (I) as well as 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl benzamide in free form has valuable pharmacological properties and can be used, for example, as an antitumor agent, as a substance for the treatment of atherosclerosis, as a substance for the treatment of restenosis, to prevent transplantation-induced disorders such as obliterative bronchiolitis and / an animal by certain bacteria such as Porphyromonas gingivalis.

Protein phosphorylation has long been known as an important step in cell differentiation and division. Phosphorylation is catalyzed by protein kinases, further divided into serine threonine kinases and tyrosine kinases. Tyrosine kinase includes Platelet-derived Growth Factor (PDGF), a receptor tyrosine kinase.

In general, PDGF is a very frequently occurring growth factor which plays an important role in both normal growth and pathological cell proliferation, as is evident in carcinogenesis and vascular smooth muscle cell disease, such as atherosclerosis and thrombosis.

In vitro inhibition of PDGF-stimulated tyrosine kinase receptor activity is measured in the PDGF receptor immune complexes of BALB / c 3T3 cells as described by E. Andrejauskas-Buchdunger and U. Regenass in Cancer Research 52, 5353-5358 (1992). The compound of formula (I), which has been described in detail, in particular its β-crystalline form, inhibits PDGF-dependent cellular receptor phosphorylation. PDGF receptor tyrosine kinase inhibition is measured by ELISA microtiter assay (see Trinks et al., J. Med. Chem., 37, 1015-1027 (1994)). 4- (4-Methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl} benzamide inhibits PDGF receptor tyrosine kinase activity at IC ₅₀ (concentration , wherein the activity is inhibited by 50% (control) at a concentration of about 120 nM and 100 nM, respectively.

By inhibiting PDGF, the compound of formula (I) is suitable for the treatment of cancer, such as gliomas, sarcomas, prostate tumors, colon, breast and ovarian tumors.

Also, abl kinase, especially v-abl kinase, is inhibited by 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl 25062003, 05:30:29 Benzamide and its methanesulfonic acid salt. Inhibition of v-abl tyrosine kinase can be determined by the methods of N. Lydon et al., Oncogene Research 5, 161-173 (1990) and JF Geissler et al., Cancer Research 52, 4492-4498 (1992). In these methods, [Val ⁵ ] -angiotensin II and [γ ^'32 ] -αΤΡ are used as substrates. 4- (4-Methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl] benzamide has an IC 50 at 38 nM.

Analogously, the salt of the compound of formula (I) also inhibits BCR-abl-kinase (see Nature Medicine 2, 561-566 (1996)) and is thus useful in the treatment of BCR-abl-positive cancer and cancer such as leukemia (especially chronic myeloid leukemia and acute lymphoblastic leukemia where active apoptotic mechanisms have been found), and also has effects on the subset of leukemic stem cells as well as the ability to purify these cells in vitro after removal of these cells (e.g. bone marrow removal) and reimplantation of these cells if clearly identified as cancer cells (e.g., reimplantation of purified bone marrow cells).

Needless to say, all of the above-mentioned inhibitory and pharmacological effects also have the 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl)] - free base pyrimidin-2-ylamino] phenyl} benzamide or other salts thereof.

The pharmacologically active compounds of the invention may be used in the form of preparations for parenteral administration or in the form of infusion solutions. Such solutions are preferably isotonic aqueous solutions or suspensions which may be prepared prior to administration, for example in the case of lyophilized preparations which contain the active substance either alone or together with a carrier, for example mannitol. The pharmaceutical substances may be sterilized and / or may contain excipients, for example, preservatives, stabilizers, wetting and / or emulsifying agents, solubilizing agents, salts for regulating osmotic pressure and / or buffers. Said pharmaceutical preparations which, if desired, may contain other pharmacologically active substances, such as antibiotics, are prepared in a known manner, for example by conventional mixing, granulating, coating, dissolving or lyophilizing, and contain from about 1% to 100%, in particular from about 1%. % to 20%, of one or more active substances.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings illustrate the invention.

Figure 1 shows an X-ray diffraction pattern of the α-crystalline form of the methanesulfonic acid addition salt of the compound of formula (I).

Figure 2 shows crystals of α-crystalline form and crystals of β-crystalline form of 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidine- 2-ylamino] phenyl} benzamide (= methanesulfonic acid addition salt of compound of formula (I)).

In the diagram, the angle of refraction 2theta is plotted on the horizontal axis (x-axis) and the relative line intensity (peak corrected by background) on the vertical axis (y-axis). The diagrams are obtained as follows: First, X-ray diffraction chart of the collected film using a Guinier camera (Enraf-Nonius FR 552 model) with a Guinier 258-94c film and the discharge between the copper electrodes (Kal radiation, wavelength λ = 1.54060 =. 1 O ¹⁰ m). The optical density of the lines on the film is proportional to the light intensity. The film is then scanned using a line scanner (LS 18, Johansson, Täby, Sweden) with SCANPI software.

Melting points are determined by a DSC thermogram using a Mettler-Toledo TA 8000. DSC (differential scanning calorimetry) is a dynamic differential calorimetry technique. Using this technique, the melting point of the α-crystalline form can be measured by heating the samples until it is not possible to detect the thermal, i. j. endothermic or exothermic reaction using ultrasensitive sensors. The melting points herein are determined using a Mettler-Toledo TA 8000 when about 5.5-6.5 mg of each sample in a perforated lid aluminum crucible, in a calm air atmosphere, is measured during heating at a rate of 10 ° C / min. . (starting from 20 ° C).

α-Crystalline form of 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl} benzamide methanesulfonate is characterized by needle-like crystals and is hygroscopic. This form does not have crystals too suitable for pharmaceutical compositions as solid dosage forms because their physical properties, such as flow characteristics, are not suitable.

The following examples illustrate the invention but do not limit it. _Rf values are determined on TLC plates coated with silica gel (Merck, Darmstadt, Germany). The ratio of solvents in the solvent systems used is expressed in volume (v / v) and temperatures are given in degrees Celsius (° C).

DETAILED DESCRIPTION OF THE INVENTION

Eluents (gradients)

HPLC gradient:

% b) in a) for 20 minutes, then 0% 30% b) in a) for 10 minutes, then 30% b) in a) for 5 minutes

Eluent a): ion pairing reagent and methanol (420 ml and 580 ml) Eluent b): ion pairing reagent and methanol (40 ml and 960 ml)

Ionic vapor reagent: 7.5 g of 1-octanesulfonic acid dissolved in about 800 ml of water, pH adjusted to 2.5 with phosphoric acid, diluted with water to 1000 ml.

Column: 150 x 3.9 mm, filled with Symmetry C18 5 µ (Waters), pre-equilibrated with eluent a). Flow rate 1.2 ml / min, UV detection at 267 nm.

Example 1

Preparation of α-crystalline form of 4- (4-methylpiperazin-1-ylmethyl) -N- (4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino) phenyl} benzamide methanesulfonate

4- (4-Methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl} benzamide methanesulfonate was prepared as follows: 98.6 g ( 0.2 mol) of free 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3-yl) pyrimidin-2-ylamino] phenyl} benzamide (for preparation see, for example, EP-A 0 564 409) is added to 1.4 L of ethanol. To this beige suspension was added dropwise 19.2 g (0.2 mol) of methanesulfonic acid over 20 minutes. The solution was refluxed for 20 minutes and then clarified by filtration at 65 ° C. The filtrate was evaporated to 50% and the residue was filtered off at 25 ° C (filtered material A). The mother liquor is evaporated to dryness. This residue and the filtered material A are suspended in 2.2 L of ethanol and dissolved under reflux with the addition of 30 ml of water. Cooling to 25 ° C overnight, filtration and drying at 65 ° C to constant weight gave 4- (4-methylpiperazin-1-ylmethyl) -N- {4-methyl- [3- (4-pyrid-3)]. -yl) pyrimidin-2-ylamino] phenyl) benzamide as light beige crystalline methanesulfonate (α-crystalline forms) ·

Claims (2)

A crystalline form of a monomethanesulfonic acid addition salt with a compound of formula (I), characterized by a melting point of about 226 ° C, defined as the onset of melting in a differential scanning calorimetry diagram. The crystalline form according to claim 1, characterized in that it has acicular crystals.